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基于环介导等温扩增法的快速可视化鉴定

Rapid and Visual Identification of With Loop-Mediated Isothermal Amplification Method.

作者信息

Wang Nan, Zhao Zhiyong, Gao Jie, Tian Enjing, Yu Wenjie, Li Hui, Zhang Juan, Xie Ruibin, Zhao Xiaoyan, Chen Ailiang

机构信息

Institute of Quality Standard and Testing Technology for Agro-Products, Key Laboratory of Agro-product Quality and Safety, Chinese Academy of Agricultural Sciences, Beijing, China.

Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, Shanghai, China.

出版信息

Front Microbiol. 2021 Mar 18;12:638315. doi: 10.3389/fmicb.2021.638315. eCollection 2021.

Abstract

is a kind of common poisonous mushroom in China that is widely distributed in different areas. Food poisoning caused by accidentally eating has become more frequent in recent years. In 2019, there were 55 food poisoning incidents caused by eating this mushroom in China. Mushroom poisoning continues to be a common health issue of global concern. When mushroom poisoning occurs, an effective, simple, and rapid detection method is required for accurate clinical treatment or forensic analysis. For the first time, we established a loop-mediated isothermal amplification (LAMP) assay for the visual detection of . A set of specific LAMP primers was designed, and the specificity was confirmed against 43 different mushroom species. The LAMP method could detect as low as 1 pg of genomic DNA. Boiled mushrooms and artificial gastric-digested mushroom samples were prepared to test the applicability of the method, and the results showed that as low as 1% in boiled and digested samples could be successfully detected. The LAMP method can also be completed within 45 min, and the reaction results could be directly observed based on a color change under daylight by the naked eye. Therefore, the LAMP assay established in this study provides an accurate, sensitive, rapid, and low-cost method for the detection of .

摘要

是中国一种常见的有毒蘑菇,广泛分布于不同地区。近年来,因误食导致的食物中毒事件愈发频繁。2019年,中国有55起因食用这种蘑菇引发的食物中毒事件。蘑菇中毒仍是一个全球关注的常见健康问题。当发生蘑菇中毒时,需要一种有效、简单且快速的检测方法来进行准确的临床治疗或法医分析。我们首次建立了一种用于可视化检测的环介导等温扩增(LAMP)检测方法。设计了一组特异性LAMP引物,并针对43种不同蘑菇物种确认了其特异性。LAMP方法能够检测低至1 pg的基因组DNA。制备了水煮蘑菇和人工胃消化蘑菇样本以测试该方法的适用性,结果表明在水煮和消化样本中低至1%的[目标物]都能被成功检测到。LAMP方法也可在45分钟内完成,并且反应结果可通过肉眼在日光下基于颜色变化直接观察到。因此,本研究建立的LAMP检测方法为[目标物]的检测提供了一种准确、灵敏、快速且低成本的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bae1/8013719/aba69616c0b8/fmicb-12-638315-g001.jpg

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